# Haynes Lab:Notebook/Engineering PC-TFs/2013/10/19

(Difference between revisions)
 Revision as of 17:20, 20 October 2013 (view source) (→October 19, 2013)← Previous diff Current revision (17:20, 20 October 2013) (view source) (→October 19, 2013) Line 12: Line 12: | align="center" style="background:#efefef;"| '''Part''' | align="center" style="background:#efefef;"| '''Part''' | align="center" style="background:#efefef;"|'''Concentration''' | align="center" style="background:#efefef;"|'''Concentration''' - - - |- |- | CMV (insert)||15.535 ng/μL | CMV (insert)||15.535 ng/μL

## Current revision

Main project page
Previous entry      Next entry

## October 19, 2013

Ligation of CMV + MV9

 Part Concentration CMV (insert) 15.535 ng/μL MV9 (vector) 20.838 ng/μL

Calculations

ng insert = bp insert/bp vector * 2 * 50
ng insert = 588/4611 * 2 *50 = 12.75211 ng

μL insert = ng insert/concentration insert
μL insert = 12.75211/15.535 = .82μL

μL vector = 50 ng/concentration vector
μL vector = 50/20.838 = 2.4 μL

 1 Control DNA Insert 0.8 0 DNA Vector (MV9) 2.4 2.4 T4 Ligase 1 1 Lign Buffer (2x) 5 5 dH2O 0.8 1.6 Total 10 µL 10 µL

Bacterial Transformation of CMV Promoter + MV9

• Warm 100 μg/mL Amp agar plate at 37°C
• Thaw fresh tube of DH5α Turbo cells on ice
• Add 30 μL of DH5α Turbo Cells to DNA + dH2O.
• This was also done for transformation of part KAH126
• Incubate cells + DNA on ice for 5 minutes.
• Label pre-warmed plate.
• Transfer cells + DNA onto agar.